Post on 14-Jan-2016
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HIV Drug Resistance Training
Module 13:
Equipment and Supplies
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Topics
Overview of Responsibilities Equipment and Supplies Needed for
Genotyping Equipment Use and Care
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Objectives
Identify specialized equipment and supply needs for a genotyping lab.
Describe how to ensure quality through appropriate care and use of equipment.
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A Systems Approach to Laboratory Quality
Organization
Stock Management
SOP, Documents & Records
Process Improvemen
t
Personnel
Quality Control
Occurrence Management
Specimen Management
Equipment
Data Management
Assessment
Safety & Waste
Management
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Functioning Equipment is Vital to Quality Service
Produces reliable
test results
Lowers repair costs
Prevents delays in
testing
Maintains productivity
QualityResults
FunctioningEquipment
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overview of responsibilities
What are the responsibilities of lab management?What are the responsibilities of lab staff/technicians?
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Management Responsibilities
In the area of… Lab mgt. must ensure…
Ordering pieces of equipment
Each piece meets needs and specifications of lab
Receiving and installing equipment
The lab is ready for installation
Accountability and supervision
Assigning oversight responsibility for equipment
Standard operating procedures
Develop/delegate and approve equipment SOPs
Operating equipment Train staff
Troubleshooting Train staff
Inventory control Establish inventory record
Maintenance Develop maintenance program
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Staff/Technician Responsibilities
Attend training and learn all procedures Properly use the instrument by following
SOPs Perform routine maintenance
– Routine function check– Troubleshooting– Maintenance log
Take corrective actions: notifying lab manager when problems arise
Keep records on maintenance and problems
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Discussion
How will you assign responsibilities for equipment among different members of management and staff?
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equipment and supplies needed for genotypingWhat equipment and supplies do all labs need for HIVDR testing?What equipment and supply decisions depend on the testing method?
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Core Laboratory Equipment
Microcentrifuge Vortex Biological safety cabinet Agarose gel electrophoresis Computer, printer, Internet
connection UV Imaging system Reliable power supply
(Uninterruptible power supply, UPS, for short outages)
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Core Laboratory Supplies
Micropipettes Agarose gel electrophoresis, power supply Refrigerators and freezers Distilled, deionized water supply Sink Ice machine Timers
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Equipment for Genotyping
DNA sequencer PCR machine Dead-air cabinet (PCR workstation) Computer attached to sequencer Specialized software dependent on Platform
used Sequencing kit
May be influenced by platform (ViroSeq,
TruGene, or In-house)
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Laboratory Supplies
Sequencer supplies: capillary array, polymer, buffers
PCR supplies: tubes, plates, trays RNA/DNA purification: columns, buffers Electrophoresis: Agarose, ethidium bromide
or other stains, buffers, MW markers/standards
Sterile, RNAse-free water Pipette tips (aerosol-resistant for pre-PCR) Polaroid film or special printing paper Disposable gloves Lab coats (disposable or not)
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Types of Pipettes
Single, multichannel, automated, repeater Use disposable, single-use, pipette tips Positive displacement and aerosol barrier
tips recommended for pre-PCR work
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Equipment: Pre-Amplification Area, In-house Assay
Essential:– Bench with sink / tap water– Biohazard flow, class IIb– Dead air cabinet (master mix preparation)– -20°C freezer and 4°C refrigerator– Microcentrifuge 12,500-15,000 g– Vortex– Timers– Dedicated set of micropipettes for master mix
and RNA extraction Optional:
– Ultracentrifuge 21,000-25,000 g– Dead air cabinet (RNA extraction)– -80°C freezer
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PCR Workstations (Dead-air Cabinets)
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Equipment: Post-Amplification Area,In-house Assay
Essential:– Bench with sink / tap water– Dead air cabinet (nested reaction)– Thermal cyclers (e.g. ABI, Bio-Rad, Hybaid, etc)– Agarose gel apparatus– Photo documentation of agarose gel– DNA sequencer (e.g. ABI, Beckman)– Computer + programs (editing)– Microcentrifuge 450-550 g– Vortex– Dedicated set of micropipettes– Timers– -20°C freezer– 4°C refrigerator
Optional:– -80°C freezer
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ABI Sequencing Instruments
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Alternative Sequencing Instruments
Beckman CEQ8000/8800
TruGene
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Remember Workflow!
Consider lab layout (module 4) when planning equipment needs and placement
LAB 1
"Specimen Processing" Lab
LAB 2
"Post-amplification" Lab
-80°C
freezer
BSC: bio-safety cabinet
Ice machine
Ice machine
-80°C
freezer
-20°C
freezer
wor
kbe
nch
Sequencer Gel electrophoresis areaPCR
machines
Lab coat rack
Lab coat rack
NA extraction equipment
PCR machines
Dead air cabinet
De
ad a
ir
cab
ine
tC
en
trif
ug
e
sink Eye wash
-20°C freezer
BSC BSC
Eye wash
Ce
ntr
ifu
ge
LAB 1
"Specimen Processing" Lab
LAB 2
"Post-amplification" Lab
-80°C
freezer-80°C
freezer
BSC: bio-safety cabinet
Ice machine
Ice machine
-80°C
freezer-80
°C
freezer-20°C
freezer-20°C
freezer
wor
kbe
nch
Sequencer Gel electrophoresis areaPCR
machines
Lab coat rack
Lab coat rack
NA extraction equipment
PCR machines
Dead air cabinet
De
ad a
ir
cab
ine
tC
en
trif
ug
e
sink Eye wash
-20°C freezer-20°C freezer
BSCBSC BSCBSC
Eye wash
Ce
ntr
ifu
ge-20°C
freezer
wor
kben
ch
Ice machine
LAB 1
"No Template" Lab
LAB 2
"Specimen Processing" Lab
LAB 4
"Post-amplification" Lab
LAB 3
"Nested PCR" Lab
-80°C
freezer
-20°C freezer
BSC
wor
kben
ch
Ice machine
Ice machine
-80°C
freezer-20°C
freezer
wor
kben
ch
Sequencer Gel electrophoresis areaPCR
machinesworkbench PCR
machines
-20°C freezer
Cen
trif
ug
e
Pass-through window
Lab coat rack
Lab coat rack
Lab coat rack
Lab coat rack
NA extraction equipment
workbench
sink
sink
sink
sinkEye
wash
Eye wash
BSC
Cen
trif
ug
e
-20°C freezer-20°C freezer
wor
kben
ch
Ice machine
LAB 1
"No Template" Lab
LAB 2
"Specimen Processing" Lab
LAB 4
"Post-amplification" Lab
LAB 3
"Nested PCR" Lab
-80°C
freezer-80°C
freezer
-20°C freezer-20°C freezer
BSCBSC
wor
kben
ch
Ice machine
Ice machine
-80°C
freezer-80°C
freezer
-20°C
freezer-20°C
freezer
wor
kben
ch
Sequencer Gel electrophoresis areaPCR
machinesworkbench PCR
machines
-20°C freezer-20°C freezer
Cen
trif
ug
e
Pass-through window
Lab coat rack
Lab coat rack
Lab coat rack
Lab coat rack
NA extraction equipment
workbench
sink
sink
sink
sinkEye
wash
Eye wash
BSCBSC
Cen
trif
ug
e
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Reflection
What is our “shopping list?”
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equipment use and care
What can we do to make sure our genotyping equipment is properly cared for?
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Maintenance
Properly maintained equipment is needed for quality results
Maintenance: Routine and biannual or yearly scheduled maintenance – Reduces breakdowns – Reduces costs in the long run– Minimizes "down time"
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Introducing New Equipment
Calibrate or verify specified performance upon installation– Pipettes– Centrifuge– Vortex– Etc.
Perform assay equivalence testing– Key components of the genotyping process
(sequencer, PCR machine)– 10-20 samples previously reported and verify
results within expected range of variability
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Maintenance: Function Checks
Verify that equipment is working properly Performed routinely
– Daily, weekly, monthly– After adjustment or repair
Example:– Monitoring refrigerator temperatures– Verifying pipette accuracy– Checking centrifuge speed
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Refrigerator and Freezer: Use and Care
Keep organized Periodically clean inside and outside Ensure door is completed sealed when
closing CAUTION! – DO NOT store food items or
beverages in laboratory refrigerator or freezer
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Refrigerator & Freezer:Temperature Checks
Monitor daily– Refrigerator: 4°C (2°C
to 8°C) – Freezer: -20°C (+/- 5°C)
or -80°C (+/- 10°C) If possible install
electronic monitoring and alarm system
Avoid "frost-free" freezers for storage of critical reagents (enzymes, specimens, etc.)
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Refrigerator & Freezer:Temperature Log
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Keep a Log for All Maintenance Activities
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Exercise: Create a Maintenance Activity List
Purpose:– Use what you have learned in this module and
create a maintenance checklist specific to your test site
Process:– Work in groups of 3-4 (or by test site) – Create a list of maintenance activities on a
daily, weekly, monthly, and yearly basis – You can use the handout as a starting point.– Activity time = 10 minutes
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Discussion
What equipment and supplies do all labs need for HIVDR testing?
What equipment and supply decisions depend on the testing method?
What challenges do you face in your setting regarding equipment and inventory supply?
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Summary
Overview of Responsibilities Equipment and Supplies Needed for
Genotyping Equipment Use and Care